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ENDOMETRIUM Publication-derived

Endometrial Organoids for Disease Modelling

Full SOP with Academic or a plan

Source Turco et al., 2017 · University of Cambridge, Department of Pathology · 10.1038/ncb3516

Authors: Margherita Y. Turco, Lucy Gardner, Jasmine Hughes, Tereza Cindrova-Davies, Maria J. Gomez, Lydia Farrell, Michael Hollinshead, Steven G.E. Marsh, Jan J. Brosens, Hilary O. Critchley, Benjamin D. Simons, Myriam Hemberger, Bon-Kyoung Koo, Ashley Moffett, Graham J. Burton Duration: 14 days 6 phases Cell source: Human endometrial epithelial cells (non-pregnant endometrium and decidua)

Abstract

This protocol establishes long-term, genetically stable 3D organoid cultures from human non-pregnant endometrium and decidua in a chemically-defined expansion medium. The organoids recapitulate endometrial glandular features in vivo, respond to sex hormones, and can differentiate into ciliated and secretory cells upon pregnancy hormone stimulation, providing a model system for studying implantation, early pregnancy, and endometrial pathologies.

Cell source
Human endometrial epithelial cells (non-pregnant endometrium and decidua)
Application
Disease modeling and developmental study of endometrial function, implantation, and early pregnancy

Protocol overview

19 steps across 6 phases

Phase 1 · Tissue Processing and Gland Isolation Day 0
  1. a Tissue chopping and enzymatic digestion
  2. b Filtration and gland collection
Phase 2 · Organoid Initiation and Plating Day 0-1
  1. a Preparation of Matrigel suspension
  2. b Plating in Matrigel droplets
  3. c Overlay with Expansion Medium (ExM)
Phase 3 · Organoid Expansion and Maintenance Day 1-14+ (passaging every 7-10 days)
  1. a Medium refreshment
  2. b Manual passaging of organoids
  3. c Long-term expansion monitoring
Phase 4 · Hormonal Differentiation – Ovarian Hormone Treatment Day 4-10
  1. a Estrogen priming
  2. b Progesterone and cAMP co-stimulation
  3. c Collection for analysis
Phase 5 · Pregnancy Signal Stimulation Day 4-12
  1. a Preparation of Differentiation Medium (DM)
  2. b Addition of pregnancy hormones
  3. c Extended differentiation culture
  4. d Collection for analysis
Phase 6 · Single-Cell Cloning and Clonogenic Assays Day 0-14
  1. a Preparation of single-cell suspension
  2. b Limiting dilution plating
  3. c Scoring organoid formation
  4. d Expansion and characterization of clonal lines

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Attribution

This SOP was written by Organthis based on the published method in Turco et al., 2017. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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